In 1996, a complete sequence of the budding yeast genome was completed and numerous studies begun to discover the function of genes. A primary concern of this study was to identify genes that may function in cytokinesis. Seven candidate genes were chosen, which were previously shown to localize to the bud neck, the site of cytokinesis, or interact with a protein that is involved in cytokinesis. Six of those uncharacterized genes, YHR149C, YLR187W, YOL070C, YMR124W, YPL158C, and YOR304C-A, were identified by data generated by GFP tagging of the yeast proteome. Data from a large-scale two-hybrid screen was used to identify YGR153W. The results from a PCR-mediated technique allowing single-step deletion of chromosomal genes revealed that those seven genes might not function in cytokinesis.
Another interest of this study was the role of phosphorylation of the PEST region of Hof1. Hof1p is a member of the pombe Cdc15p homology (PCH) family of proteins that localizes to actin-rich regions, such as cytokinetic actin ring. To determine if phosphorylation of the PEST region of Hof1 was important to regulate Hof1 dynamics and cytokinesis, in vitro mutagenesis on potential phosphorylation sites of the PEST domain was performed and further characterization is required. --Abstract, page iii
